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Cut&Run of female mouse primordial germ cells at E11.5 and E12.5 days of gestation for H3K27me3

ABSTRACT: To evaluate the impacts of X-chromosome reactivation on repressive H3K27me3 chromatin marks, allele-specific low input CUT&Run experiments were conducted starting from genomic DNA obtained from FACS-sorted primordial germ cells.

ORGANISM(S): Mus musculus

PROVIDER: GSE243464 | GEO | 2025/01/21

REPOSITORIES: GEO

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Cut&Run of female mouse primordial germ cells at E11.5 and E12.5 days of gestation for H3K27me3

Project description:Cut&Run of female mouse primordial germ cells at E11.5 and E12.5 days of gestation for H3K27me3

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SLIC-CAGE (Super-Low Input Carrier-CAGE) application on mouse primordial germ cells isolated from embryonic day E11.5

Project description:We developed SLIC-CAGE (Super-Low Input Carrier-CAGE) approach to capture 5'end of RNA polymerase II transcripts from as little as 5-10 ng of total RNA. The dramatic increase in sensitivity compared to existing CAGE methods is achieved by specially designed, selectively degradable carrier RNA. We apply SLIC-CAGE on mouse primordial germ cells embryonic day (E) 11.5 - 2 biological replicates.

2018-10-25 | E-MTAB-7056 | biostudies-arrayexpress

Chicken embryonic stem cell/primordial germ cell LC-MSMS

Project description:As germ cell precursor, primordial germ cells (PGCs) are widely used in transgenic animal production, regenerative medicine and other fields. However, the regulation mechanism of chicken PGCs is not incomplete, which leads to the insufficient amount of chicken PGCs obtained in vitro, which seriously affects the specific application of PGCs. During PGC formation (differentiation from ESCs to PGCs), some proteins have inconsistent changes in transcription level and protein abundance. Mediating proteasome degradation is one of the most important roles of protein ubiquitination, and enrichment analysis of transcriptome and proteome also suggests an important role of ubiquitination in the process of PGCs. In order to explore the important functions and potential targets of ubiquitination, we collected chicken ESCs and PGCs cells for label free ubiquitomics analysis. This study preliminarily analyzed how ubiquitination regulates the formation of chicken PGCs, providing a theoretical basis for the subsequent research and specific application of PGCs.

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In vitro modeling of primordial germ cell development

2009-06-22 | GSE7948 | GEO

PRDM14 controls X-chromosomal and global epigenetic reprogramming of H3K27me3 in migrating mouse primordial germ cells.

Project description:BackgroundIn order to prepare the genome for gametogenesis, primordial germ cells (PGCs) undergo extensive epigenetic reprogramming during migration toward the gonads in mammalian embryos. This includes changes on a genome-wide scale and additionally in females the remodeling of the inactive X-chromosome to enable X-chromosome reactivation (XCR). However, if global remodeling and X-chromosomal remodeling are related, how they occur in PGCs in vivo in relation to their migration progress and which factors are important are unknown.ResultsHere we identify the germ cell determinant PR-domain containing protein 14 (PRDM14) as the first known factor that is instrumental for both global reprogramming and X-chromosomal reprogramming in migrating mouse PGCs. We find that global upregulation of the repressive histone H3 lysine 27 trimethylation (H3K27me3) mark is PRDM14 dosage dependent in PGCs of both sexes. When focusing on XCR, we observed that PRDM14 is required for removal of H3K27me3 from the inactive X-chromosome, which, in contrast to global upregulation, takes place progressively along the PGC migration path. Furthermore, we show that global and X-chromosomal reprogramming of H3K27me3 are functionally separable, despite their common regulation by PRDM14.ConclusionsIn summary, here we provide new insight and spatiotemporal resolution to the progression and regulation of epigenome remodeling along mouse PGC migration in vivo and link epigenetic reprogramming to its developmental context.

| S-EPMC6585054 | biostudies-literature

Role of Ror2 in primordial germ cell migration

Project description:Primordial germ cells (PGCs), the embryonic precursors of eggs and sperm, are a unique model for identifying and studying regulatory mechanisms in singly migrating cells. From their time of specification to eventual colonization of the gonad, mouse PGCs traverse through and interact with many different cell types, including epithelial cells and mesenchymal tissues. Work in drosophila and zebrafish have identified many genes and signaling pathways involved in PGC migration, but little is known about this process in mammals. We have generated a point mutation in the Ror2 gene that we know disrupts primordial germ cell migration in the developing mouse embryo. We used microarray analysis to determine if this defect is mediated through genome-wide or pathway-specific transcriptional changes. We analyzed primordial germ cells (PGCs) from 4 wild-type (WT) and 4 Ror2Y324C/Y324C mutant embryos using Oct4-DPE-EGFP. PGCs were collected during their active migratory state at embryonic day 9.5 (somite range 20-25).

2014-08-08 | E-GEOD-60179 | biostudies-arrayexpress

Transcription profiling of mouse primordial germ cells in vitro modeling of primordial germ cell development

Project description:Transgenic StellaGFP ESCs were used to derive primordial germ cells during embryoid body (EB) differentiation, and microarry analysis used to compared FACS sorted Stella-positive cells of day 7 Ebs with the parental ESCs and Stella-negative cells of day 7 Ebs. Experiment Overall Design: Four replicates of Stella-positive day 7 cells were compared to two replicates of stella-negative day 7 cells. Also, there were three replicated of FACS sorted Stella-negative ESCs, two replicated of FACS-sorted Stella-postive ESCs, and two replicates of unsorted StellaGFP ESCs.

2009-07-02 | E-GEOD-7948 | biostudies-arrayexpress

Role of Dppa3 during primordial germ cell development I

Project description:We performed whole-genome bisulfite sequencing and RNA-seq analysis of Stella knockout primordial germ cells (PGCs).

2023-04-27 | GSE196620 | GEO

Offspring from oocytes derived from in vitro primordial germ cell-like cells in mice

Project description:Reconstitution of female germ-cell development in vitro is a key challenge in reproductive biology and medicine. We show here that female (XX) embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) in mice are induced into primordial germ cell-like cells (PGCLCs), which, when aggregated with female gonadal somatic cells as reconstituted ovaries, develop pre-meiotic germ cell characteristics, including X-reactivation, imprint erasure, and cyst formation. Upon transplantation under ovarian bursa, PGCLCs in the reconstituted ovaries mature into germinal vesicle-stage oocytes, which, through in vitro maturation and fertilization, contribute to fertile offspring. Our culture system serves as a robust foundation for the investigation of key properties of female germ cells, including the acquisition of totipotency, and for the reconstitution of whole female germ-cell development in vitro.

2012-11-05 | GSE40716 | GEO

Proteome landscape of mouse primordial germ cells

Project description:Primordial germ cells (PGCs) are precursors of both male and female gametes as fundamental materials for organism development. The transcriptome, methylome and chromatin accessibility profiles of PGCs in both mice and humans have been recently reported. However, little is known about the characteristics of PGCs at the protein levels, which directly exert cellular functions. Here, we construct landscapes of both proteome of mouse PGCs at E11.5, E13.5 and E16.5 days, the three critical developmental windows for PGCs’ sex differentiation, female meiosis initiation and male mitotic arrest. In each developmental stage of PGCs nearly 2000-3000 proteins are identified, among which specific functional pathways such as oxidative phosphorylation, DNA damage repair, and meiotic cell cycle are involved for different events during PGCs development. Thus, our work offers a valuable resource for the systematic investigations of PGC development at protein level.

| PXD019222 |

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